PROJECT SUMMARY The Proteomics Core (PC) provides state-of-the-art protein chemistry services for Moffitt Cancer Center (MCC) members and cutting-edge analytical techniques to qualitatively and quantitatively assess protein expression, interactions, activity, mutations and post-translational modifications (PTMs). The core was established in 2003 and was scored ?Outstanding? in the 2011 review. Experiments can be conducted with cell line and animal models as well as with patient specimens, including diverse tissue types and biofluids. The scope of these studies ranges from detailed molecular characterization of a single protein to proteome-wide profiling. The overall aims of the Proteomics Core are to: 1) consult and collaborate with members in the design and execution of proteomics experiments; 2) provide instrumentation and highly trained staff to support members; and 3) train members and their staff. The PC is comprised of four full-time staff and provides liquid chromatography-tandem mass spectrometry (LC-MS/MS) peptide sequencing for discovery proteomics, and liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM) for targeted quantification. Studies performed in the PC include those that examine protein-protein interactions (using immunoprecipitation or tandem affinity purification), drug targets (chemical proteomics), signaling (phosphoproteomics), proteome- wide acetylation or ubiquitination, sequential enrichment of PTMs, and the effects of different perturbations on selected proteins or the proteome (e.g., drug treatment, siRNA knockdown, CRISPR knockout). Further, the PC provides sample multiplexing using metabolic labeling (SILAC) or chemical labeling (iTRAQ/TMT) techniques for comparative proteomics. Finally, the PC uses an integrated pipeline of discovery proteomics to explore cancer biology and target quantification for highly precise biomarker measurements that can be translated from model systems into patient specimens. PC interactions with the Molecular Genomics Core (MGC) are necessary for combined proteogenomic analysis; and with the University of Florida South East Center for Integrated Metabolomics, to support proteometabolomics. Downstream interactions with the Biostatistics Core (BC) and the Cancer Informatics Core (CIC) contribute to understanding and evaluating the results of complex proteomics experiments. During the project period, the PC supported users across four and contributed to 47 publications, an increase from 15 publications in the 2006-2010 funding period. In the most recent fiscal year, the PC supported 26 members, with 90% of total usage by peer-review-funded members. Using financial support provided by the CCSG and Institutional funds, the PC continues to provide state-of-the- art equipment, technologies, and services to meet the current and future needs of MCC members.